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外源性硫化氢对大鼠移植肺细胞凋亡的影响
Effect of exogenous hydrogen sulfide on apoptosis of pneumonocytes in rats after lung transplantation

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DOI:
作者:
陈旭，吴镜湘，朱宏伟，徐美英
Chen Xu,Wu Jingxiang,Zhu Hongwei,Xu Meiying
作者单位:
上海交通大学附属胸科医院麻醉科
Department of Anesthesia, Shanghai Chest Hospital, Shanghai Jiaotong University ,Shanghai, China, 200030
关键词:
硫化氢；肺移植； 肺缺血再灌注损伤；凋亡； Bax； Bcl-2
H2S; lung transplantation; Lung Ischemia Reperfusion Injury; apoptosis; Bax;Bcl-2
摘要:
目的 探讨外源性H2S对大鼠移植肺细胞凋亡的影响。方法 以硫氢化钠(NaHS) 作为外源性H2S供体，30只Sprague-Dawley(SD)雄性大鼠随机分为3组（n＝6）。假手术组（sham组）：开胸游离左肺门，未行肺移植；肺移植组（LTx组）：左肺移植后开放再灌注2h；NaHS干预组（H2S组），在再灌注开放前1h腹腔注射NaHS（14μmol•kg-1）。再灌注2h后取移植肺组织，应用脱氧核苷酸末端转移酶介导的生物素化dUTP 缺口末端标记技术(TUNEL) 检 测 细 胞 凋 亡并计算凋亡指数（apoptosis index, AI）, 采用实时逆转录-聚合酶链反应( RT-PCR) 检测各组肺组织中凋亡相关基因Bax及Bcl-2的mRNA 表 达并计算Bax/Bcl-2比值。结果 LTx组与sham组相比, 凋亡指数(AI) 明显升高(P <0.05), 凋亡相关基因Bax/Bcl-2比值显著升高(P <0.05)；H2S 组与LTx组相比，AI明显降低(P <0.05)，Bax/Bcl-2比值显著降低(P <0.05)。结论 外源性H2S减少了大鼠移植肺缺血再灌注后的细胞凋亡，这可能是H2S在大鼠肺移植后肺缺血再灌注损伤中发挥保护性作用的机制之一。
【Abstract】Objective To explore the effect of exogenous hydrogen sulfide on apoptosis of pneumonocytes in rats after lung transplantation(LTx). Methods Thirty male Sprague - Dawley rats were randomly divided into three groups (n = 6). We performed single left LTx in male Sprague-Dawley rats after 3 h of cold ischemia time in LTx group. In H2S group, H2S donor NaHS (14 μmol/kg, intraperitoneally) was administered 1 h before the start of reperfusion. The sham operation group received an equivalent volume of saline. The apoptosis of pneumonocytes was estimated at 2 h after reperfusion by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling technique (TUNEL). In addition, apoptosis regulatory genes Bax (pro-apoptosis) and Bcl-2 (anti-apoptosis) were further estimated by Real-time RT-PCR analysis. Results In the lung-transplanted rats, the number of TUNEL positive cells was significantly increased (P < 0.05). Increased expression ratio of Bcl-2/Bax is associated with referfusion-mediated apoptosis in rat lungs (P < 0.05). However, NaHS administration clearly reduced apoptosis of pneumonocytes in rats after LTx, with the ratio of Bcl-2/Bax decreasing (P < 0.05). Conclusion Exogenous H2S was associated with a significant decrease in the presence of apoptosis after LTx, a suspected mediator of ischemia-reperfusion injury, which may be one of the mechanism involved with its cytoprotection against LIRI.

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